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Author:
Date:
2013-08-05
[Abstract] Transfection of primary T cells can be challenging. This protocol describes a method to transfect primary human naive CD4+ T cells with an AP-1 luciferase reporter using low-level activation by phytohemagglutinin (PHA) and electroporation, as published (Palin et al., 2013). This technique is a modification of one previously described by our group (Cron et al., 2013). Anyone wishing to transfect murine T cells should consult the publication by Cron et al., 2013. This technique may be adapted for other primary T cell types by optimizing the Neon electroporation conditions, as described in the text. Other luciferase or GFP reporters may be used, and will require optimization of the stimulation conditions for that particular reporter.
[摘要] 原代T细胞的转染可能是具有挑战性的。 该方案描述了使用植物凝集素（PHA）和电穿孔的低水平激活，用AP-1荧光素酶报道子转染初级人初始CD4 + T细胞的方法，如公开的（Palin等人 。，2013）。 这种技术是先前由我们组描述的技术的修改（Cron等人，2013）。 任何希望转染鼠T细胞的人都应参考Cron等人2013年的出版物。该技术可以通过优化氖电穿孔条件来适应其它原代T细胞类型，如文中所述。 可以使用其他荧光素酶或GFP报道分子，并且将需要优化该特定报道分子的刺激条件。
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